BrdU anticorps (Bromodeoxyuridine)

Details for Product anti-BrDU Antibody No. ABIN2477713
Antigène
Reactivité
Chemical
144
64
31
5
5
4
1
1
1
Hôte
Souris
238
35
24
19
2
1
Clonalité (Clone)
Monoclonal ()
Conjugué
Cet anticorp BrdU est non-conjugé
21
9
6
5
4
4
4
4
4
4
3
3
3
3
3
1
1
1
Application
Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
143
134
119
107
39
35
29
26
24
15
10
2
2
2
2
1
1
1
Options
'Independent Validation' signe
Antigène BrdU
Numéro du lot 1710
Application validée Immunohistochemistry
Contrôle positif

Adult (24 months) mouse brain and intestine (duodenum)

Contrôle négative

We incubated slices overnight with the blocking solution only and then processed them with the secondary antibody.

Conclusion

Passed. ABIN2477713 works well in IHC-P, but with higher concentrations than the ones suggested by the manufacturer.

Anticorps primaire ABIN2477713
Anticorps secondaire goat anti-mouse AF488 conjugated antibody (Thermo Fisher Scientific, A11029, lot 1942237)
Protocole
  • Inject mouse intraperitoneally at the dose of 0.1 mg/g body weight with BrdU (Sigma, B5002) dissolved in sterile distilled water at final concentration of 100mg/ml.
  • Perfuse mouse was perfused with paraformaldehyde 4% in 0.1M phosphate buffer (PB) pH 7.4 and post-fix samples in the same fixative for an additional 2h at RT.
  • Wash, dehydrate, and embed samples in paraffin wax.
  • Following several washes in PBS, cut intestines and brain with a microtome (6µm-thick sections) and mount sections on glass slides.
  • After paraffin removal, process sections by microwave antigen retrieval for 10min (95-100°C) in 10mM sodium citrate buffer (pH6.0).
  • Let sections cool for 20min.
  • Wash sections 2x 5min with distilled water.
  • Wash sections 5min with PBS.
  • Incubate sections in PBS containing 1% albumin from chicken egg white (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot 00583) for 1h at RT to block non-specific binding sites.
  • Incubate sections with primary antibody mouse anti-BrdU (antibodies-online, ABIN2477713, lot 1710) diluted 1:10 in PBS-BSA-PLL ON at RT.
  • Wash sections 3x 5min with 0.01M PBS.
  • Incubate sections with the secondary goat anti-mouse AF488 conjugated antibody (Thermo Fisher Scientific, A11029, lot 1942237) diluted 1:200 in 0.1M PBS for 1h at RT.
  • Wash sections 3x 5min with 0.01M PBS.
  • Mount sections in Fluoroshield (Sigma, F6182, lot MKCB0153V).
  • Acquire images with Leica DM 6000B fluorescence microscope equipped with the manufacturer’s FITC filter set and digital camera at magnifications of 20x and 40x. Parameters for image acquisition (Exposure 2.2s, Gain 2.3x, Saturation 1.5, Gamma 1.10) were maintained unchanged for all images.
Notes

For the incubation with the primary antibody ABIN2477713 1:25, 1:20, 1:15 dilutions were also tested with also 0.01M PBS - 5% Normal Goat Serum (NGS Sigma, G9023, lot SLBV1396) and 0.1% Triton-X -100 as the blocking solution, but 1:10 and PBS containing 1% albumin from chicken egg white and 0.3% Triton-X gave the best results.

Validation Images
Immunohistochemistry validation image for anti-Bromodeoxyuridine (BrDU) antibody (ABIN2477713) Staining of mouse brain and intestine sections using ABIN2477713. A-C: A bright stain...
'Independent Validation' signe
Antigène BrdU
Numéro du lot 1807
Application validée Immunohistochemistry
Contrôle positif

Adult (24 months) mouse brain and intestine (duodenum)

Contrôle négative

We incubated slices overnight with the blocking solution only and then processed them with the secondary antibody.

Conclusion

Passed. ABIN2477713 works well in IHC-P on murine intestinal samples at a 1:10 dilution.

Anticorps primaire ABIN2477713
Anticorps secondaire goat anti-mouse AF488 conjugated antibody (Thermo Fisher Scientific, A11029, lot 1942237)
Protocole
  • Inject mouse intraperitoneally at the dose of 0.1mg/g body weight with BrdU (Sigma, B5002) dissolved in sterile distilled water at final concentration of 100mg/ml.
  • Perfuse mouse with 4% paraformaldehyde in 0.1M phosphate buffer (PB) pH7.4 and post-fix samples in the same fixative for an additional 2h at RT.
  • Wash, dehydrate, and embed samples in paraffin wax.
  • Following several washes in PBS, cut intestines and brain with a microtome (6µm-thick sections) and mount sections on glass slides.
  • After paraffin removal, process sections by microwave antigen retrieval for 10min (95-100°C) in 10mM sodium citrate buffer pH6.0.
  • Let sections cool for 20min.
  • Wash sections 2x 5min with distilled water.
  • Wash sections 5min with PBS.
  • Block sections in PBS containing 1% albumin from chicken egg white (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot 00583) for 1h at RT to block non-specific binding sites.
  • Incubate sections with primary mouse anti-BrdU antibody (antibodies-online, ABIN2477713, lot 1807) diluted 1:10 in PBS-BSA-PLL ON at RT.
  • Wash sections 3x 5min with 0.01M PBS.
  • Incubate sections with secondary goat anti-mouse AF488-conjugated antibody (Invitrogen, A11029, lot 1942237) for 1h at RT.
  • Wash sections 3x 5min with 0.01M PBS.
  • Mount sections in Fluoroshield (Sigma, F6182, lot MKCB0153V).
  • Acquire images with Leica DM 6000B fluorescence microscope equipped with the manufacturer’s FITC filter set and digital camera at magnifications of 20x and 40x. Maintain parameters for image acquisition (Exposure 2.2s, Gain 2.3x, Saturation 1.5, Gamma 1.10) unchanged for all images.
Notes

For the incubation with the primary antibody ABIN2477713 dilutions 1:25, 1:20, and 1:15 were also tested, but 1:10 gave the best results.

Validation Images
Immunohistochemistry validation image for anti-Bromodeoxyuridine (BrDU) antibody (ABIN2477713) Staining in the mouse intestine (duodenum) with mouse anti-BrdU antibody ABIN2477713.
Immunogène Bromodeoxyuridine conjugated to BSA
Clone Bu20a
Isotype IgG1
Attributs du produit Mouse anti BrdU antibody, clone Bu20a, recognizes BrdU and other thymidine analogs; 5′-chloro-2′-deoxyuridine (CldU), 5′-iodo-2′-deoxyuridine (IdU) and 2′-deoxy-5-ethynyluridine (EdU), but only shows minimal reactivity with thymidine itself
Purification Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
Antigène
Autre désignation BrdU
Classe de substances Chemical
Sujet Synonyms: 5-BROMODEOXYURIDINE
Indications d'application Flow: 1/25-1/100, ICC 1/25-1/100
For Flow Cytometry use 10 μl of the suggested working dilution to label 1x106 cells in 100 μl
Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL
Buffer Phosphate buffered saline, 0.09%Sodium Azide (NaN3)
Agent conservateur Sodium azide
Précaution d'utilisation This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
Conseil sur la manipulation This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Stock 4 °C/-20 °C
Stockage commentaire Store at +4oC or at -20oC if preferred. Storage in frost-free freezers is not recommended.
Date de péremption 18 months
Images (Fournisseur)
Flow Cytometry (FACS) image for anti-Bromodeoxyuridine (BrDU) antibody (ABIN2477713) anti-Bromodeoxyuridine (BrDU) antibody
Produit citée dans: Innis, Dai, Wu, Buchan, Jacobson: "Perinatal lipid nutrition alters early intestinal development and programs the response to experimental colitis in young adult rats." dans: American journal of physiology. Gastrointestinal and liver physiology, Vol. 299, Issue 6, pp. G1376-85, 2010 (PubMed).

Background publications Dyson: "Lameness due to pain associated with the distal interphalangeal joint: 45 cases." dans: Equine veterinary journal, Vol. 23, Issue 2, pp. 128-35, 1991 (PubMed).

Magaud, Sargent, Clarke, Ffrench, Rimokh, Mason: "Double immunocytochemical labeling of cell and tissue samples with monoclonal anti-bromodeoxyuridine." dans: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, Vol. 37, Issue 10, pp. 1517-27, 1989 (PubMed).

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